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com
580

Effect of Different Stocking Densities on Hematological and Biochemical Parameters of Silver Carp,
Hypophthalmichthys molitrix Fingerlings

Salah M. Kamal
1
and Wael A. Omar
2*


1
Department of Aquaculture, Central Laboratory for Aquaculture Research at Abbassa, Agricultural
Research Centre, Egypt
2
Department of Zoology, Faculty of Science, Cairo University, Egypt
salahmk62@yahoo.com
*
ecowael@yahoo.com

Abstract: The impact of rearing silver carp, Hypophthalmichthys molitrix to the fingerling stage under three
different stocking densities was investigated depending on the hematological and biochemical parameters as
indicators of general health state of fish. The present study was carried out for 12 weeks to determine the most
optimum stocking density for rearing silver carp. Fish were cultured in duplicates of cement ponds under stocking
densities of 3, 6 and 9 fish/m
3
as T
1
, T
2
and T
3
respectively. The ponds were fertilized weekly with organic fertilizer
at rate of 50 g/m
3
. Results of hematological analyses showed significant increase in T
1
for values of RBCs count,
hemoglobin, hematocrit and mean corpuscle volume (MCV) while the lowest values were recorded in T
3
for all
these parameters. Changing the stocking density had non-significant effect on values of WBCs count, mean
corpuscle hemoglobin (MCH) and mean corpuscle hemoglobin concentration (MCHC). Plasma biochemical
analyses showed that increasing the stocking density caused significant increase in values of plasma glucose, total
protein, albumin (A) and globulin (G) associated with significant decrease in values of cholesterol and triglycerides.
The highest values of A/G ratio were recorded non-significantly in T
1
and T
3
while the lowest values were recorded
significantly in T
2
. Meanwhile T
1
showed a significant decrease in values of alkaline phosphatase (ALP), aspartate
aminotransferase (AST), alanine aminotransferase (ALT) and uric acid. Values of creatinine showed non-significant
increase among treatments. The findings suggest that the most optimum condition was detected in T
1
(3 fish/m
3
) where
most of the studied hematological and biochemical parameters were essentially normal and within the range consistent
with good fish health.
[Salah M. Kamal

and Wael A. Omar Effect of Different Stocking Densities on Hematological and Biochemical
Parameters of Silver Carp, Hypophthalmichthys molitrix Fingerlings] Life Science Journal, 2011; 8(4):580-586]
(ISSN: 1097-8135). http://www.lifesciencesite.com.

Key words: Silver carp, Hypophthalmichthys molitrix, Stocking density, Hematology, Biochemical parameters.

1. Introduction:
Aquaculture is currently the largest single
source of fish supply in Egypt accounting for almost
65% of the total fish production of the country. Most
aquaculture production in Egypt depends on
freshwater or brackish water species in the Nile delta
region with tilapias, mullets and carps making up
more than 97% of the total country production in
2007 (GFCM, 2010). Silver carp,
Hypophthalmichthys molitrix is a native species in
China and Eastern Siberia but has been intentionally
distributed all over the world for aquaculture and to
control algal blooms. It is not only utilized as human
food but also appreciated by its ability to clean water
reservoirs from clogging algae (FAO, 2005). Carps
have advantageous biological characteristics such as
wide tolerance to environmental conditions, high
growth rate and efficient filter feeding (Shen, 2009).
Silver carp is a typical planktonivore that
consumes diatoms, dinoflagellates, chrysophytes,
xanthophytes, some green algae and cyanobacteria.
In addition, detritus, conglomerations of bacteria,
rotifers and small crustaceans constitute major
components of their natural diet. Generally, there is
no need to provide supplementary formulated diet in
silver carp culture which makes its production costs
lower than most other cultured species (FAO, 2005).
The researches devoted to the evaluation of
some biochemical parameters in cultured fish are
justified by their significance for estimating the
general health condition of the animals, as well as the
possible effects of food availability, stocking density
and exposure to stressors (Kebus et al., 1992; Roche
and Boge, 1996; Chen et al., 2004; Tintos et al.,
2006). Some investigations have evidenced important
modifications of such indices under over-density
stress in cultured cyprinids (Misaila et al., 2005).
Many studies were concerned with evaluation
of hematological and biochemical parameters of
carps in normal conditions or when exposed to
stressors (Koedprang et al., 2002; Kopp et al., 2009;
Nicula et al., 2010). Most of these studies were
conducted on more than 3 years old carps but
relatively few information in the literature has
reported these parameters in carp fry or fingerlings
such as the work of Kopp et al. (2011) although it is
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581

commonly known that modification of fish blood
constituents during these early stages of development
could affect the fish performance in the future.
The aim of the present work was to evaluate the
effects of different stocking densities on
hematological and biochemical parameters of silver
carp, H. molitrix fingerlings reared in cement ponds
under normal environmental conditions.

2. Materials and Methods:
The present study was conducted during
summer season, 2009 for 12 weeks at the Research
Unit of Central Laboratory for Aquaculture Research
(CLAR), Abbassa, Abo-Hammad, Sharkia
Governorate, Egypt.

Experimental design:
A total number of 2160 silver carp with mean
body weight of 1.2 0.2 g/fish were divided into
three treatments with stocking densities of 3, 6 and 9
fish/m
3
as T
1
, T
2
and T
3
respectively. Fish were
randomly distributed at the desired stocking density
and acclimated for two weeks in duplicates of cement
ponds each measuring 20 x 2.5 x 1.2m (length x
width x water depth). The total volume of each pond
was 60 m
3
. Before starting the experiment all fish
ponds were drained completely and then were
exposed to solar radiation for 2 weeks till complete
dryness. Ponds were then refilled with freshwater
from Ismailia Canal (branched from the River Nile).
Fish ponds were fertilized weekly with Rigirs
at rate of 50 g/m
3
. Rigirs is an organic fertilizer
produced by Misr El-Salam International Company
for producing organic fertilizers. Rigirs consist of
compressed and pelleted chicken manure also heat
treated in order to be free from parasites, Salmonella,
Shigella and E. coli.

Sampling and laboratory methods:
At the end of experiment, blood was collected
from the caudal vein of 8 fish per treatment using
sodium citrate as anticoagulant and examined directly
for total red blood cells (RBCs) and total white blood
cells (WBCs) using improved Neubauer
Hemocytometer and expressed as the number of
cells/mm
3
. Hematocrit (Ht) was determined through
microhematocrit tubes using hematocrit centrifuge
for 5 min. Hemoglobin concentration (Hb) was
estimated using cyanomethemoglobin method
(Blaxhall and Daisley, 1973). The red cell indices;
mean corpuscle volume (MCV), mean corpuscle
hemoglobin (MCH) and mean corpuscle hemoglobin
concentration (MCHC) were calculated using
standard formulae (Coles, 1986). Blood samples were
centrifuged to get plasma for the determination of
glucose (Trinder, 1969), cholesterol (Allain et al.,
1974), triglycerides (McGowan et al., 1983), total
protein (Bradford, 1976), albumin content (Dumas
and Biggs, 1972), alkaline phosphatase activity
(ALP, EC. 3.1.3.1) (Bergmeyer, 1974), aspartate
aminotransferase activity (AST, EC. 2.6.1.1), alanine
aminotransferase activity (ALT, EC. 2.6.1.2)
(Reitman and Frankel, 1957), creatinine (Henry et al.,
1974) and uric acid (Barham and Trinder, 1972)
using enzymatic-colorimetric methods by means of
commercial kits (Biodiagnostic Co, Egypt). The
globulin content (G) was estimated by subtracting the
albumin content (A) from total protein content then
A/G ratio was calculated.

Statistical analysis:
The results were expressed as mean S.E. Data
were statistically analyzed using one-way analyses of
variance (ANOVA) and significance was expressed
using F-value at P<0.05. Duncans multiple range
test was used to evaluate the comparison between
means as indicated by different case letters in a
descending order A, B and C using Statistical
Analysis System, version 9.1 (SAS, 2006).

3. Results:
During the experimental period all treatments
showed non-significant mortality with over than 83%
survival in all groups also water quality parameters
remained well within limits recommended for
freshwater fish culture. Results of hematological
analyses are presented in Table 1. Statistical analysis
showed significant decrease (P<0.05) in values of
RBCs with the highest values recorded in T
1
and the
lowest values were recorded non-significantly
(Duncans test) in T
2
and T
3
. Meanwhile, changing
the stocking density had non-significant effect on
values of WBCs, MCH and MCHC. Values of Hb
and Ht showed highly significant decrease (P<0.01)
among treatments with the highest values recorded in
T
1
and the lowest ones recorded in T
3
for both
parameters. Values of MCV showed significant
decrease (P<0.05) among treatments with
significantly lower values recorded in T
3
.
As indicated in Table 2, the increase in
stocking density was associated with highly
significant increase (P<0.01) in plasma glucose and
highly significant decrease (P<0.01) in plasma
cholesterol and triglycerides. The highest values of
cholesterol were recorded non-significantly in T
1
and
T
2
while those of triglycerides were recorded
significantly in T
1
.
Results of the plasma protein content (Table 3)
showed highly significant increase (P<0.01) in values
of total protein, albumin and globulin associated with
the increase in stocking density. Meanwhile, values
of A/G ratio showed significant difference among
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treatments (P<0.05) with the highest values recorded
non-significantly in T
1
and T
3
and the lowest values
recorded significantly in T
2
.
In case of plasma indicative parameters of liver
functions (ALP, AST and ALT) (Table 4), values of
ALP, AST and ALT showed highly significant
increase (P<0.01) with the lowest values recorded
significantly in T
1
for ALP and AST. Meanwhile, the
lowest values were recorded non-significantly in T
1

and T
2
for ALT. The measured plasma creatinine and
uric acid (Table 4) were used as indicators of fish
kidney functions. Values of creatinine showed non-
significant increase among treatments. Meanwhile,
values of uric acid showed highly significant increase
(P<0.01) with the highest values recorded non-
significantly in T
2
and T
3
while the lowest values
were recorded significantly in T
1
.

Table 1: Effect of different stocking densities on hematological indices of silver carp
RBCs
(X10
6
cells/mm
3
)
WBCs
(X10
3
cells/mm
3
)
Hb
(g/dl)
Ht
(%)
MCV
(m
3
)
MCH
(pg)
MCHC
(%)
T
1
(3 fish/m
3
)
1.04 0.01
A
21.33 1.76
A
2.97 0.09
A
16.32 1.68
A
158.61
16.63
A
28.62 0.85
A
18.66 4.51
A
T
2
(6 fish/m
3
)
0.68 0.11
B
23.33 4.67
A
1.87 0.07
B
8.35 1.17
B
149.73
25.42
A
29.94 2.78
A
23.0 5.09
A
T
3
(9 fish/m
3
)
0.64 0.08
B
17.33 2.91
A
1.57 0.09
C
4.07 0.46
C
81.44 12.57
B
24.15 3.14
A
38.3112.5
A
F-value 8.30* 0.84 81.50** 32.19** 5.96* 1.51 1.58
Data are represented as means of eight samples S.E.
Means with the same letter in the same column for each parameter are not significantly different.
* Significant difference (P<0.05) ** Highly significant difference (P<0.01)

Table 2: Effect of different stocking densities on plasma glucose, cholesterol and triglycerides of silver carp

Glucose
(mg/dl)
Cholesterol
(mg/dl)
Triglycerides
(mg/dl)
T
1
(3 fish/m
3
)
72.90 7.43
C
174.09 3.06
A
181.43 6.99
A
T
2
(6 fish/m
3
)
97.52 5.32
B
150.01 2.55
A
101.79 29.23
B
T
3
(9 fish/m
3
)
102.33 10.21
A
57.72 5.63
B
64.17 3.50
B
F-value 13.94** 12.63** 11.75**
Data are represented as means of eight samples S.E.
Means with the same letter in the same column for each parameter are not significantly different.
* Significant difference (P<0.05) ** Highly significant difference (P<0.01)

Table 3: Effect of different stocking densities on plasma protein content of silver carp
Total protein
(g/dl)
Albumin (A)
(g/dl)
Globulin (G)
(g/dl)
A/G ratio
T
1
(3 fish/m
3
)
3.08 0.27
B
1.75 0.18
B
1.33 0.13
C
1.32 0.13
A
T
2
(6 fish/m
3
)
4.91 0.16
A
2.22 0.19
A
2.69 0.10
A
0.83 0.10
B
T
3
(9 fish/m
3
)
5.54 0.15
A
3.39 0.28
A
2.15 0.13
B
1.58 0.23
A
F-value 36.23** 15.03** 33.88** 5.79*
Data are represented as means of eight samples S.E.
Means with the same letter in the same column for each parameter are not significantly different.
* Significant difference (P<0.05) ** Highly significant difference (P<0.01)


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583

Table 4: Effect of different stocking densities on indicative parameters of liver and kidney functions in
plasma of silver carp

ALP
(U/l)
AST
(U/l)
ALT
(U/l)
Creatinine
(mg/dl)
Uric acid
(mg/dl)
T
1
(3 fish/m
3
)
53.32 1.92
B
16.63 4.28
C
14.88 3.17
B
0.28 0.12
A
2.21 0.19
B
T
2
(6 fish/m
3
)
82.26 4.70
A
21.75 4.96
B
15.88 1.30
B
0.31 0.08
A
3.15 0.40
A
T
3
(9 fish/m
3
)
83.76 5.70
A
25.13 6.89
A
23.25 2.98
A
0.35 0.10
A
2.82 0.43
A
F-value 12.72** 13.61** 10.61** 0.68 11.82**
Data are represented as means of eight samples S.E.
Means with the same letter in the same column for each parameter are not significantly different.
* Significant difference (P<0.05) ** Highly significant difference (P<0.01)

4. Discussion:
Fish production capacity can increase by
identifying environmental factors and providing
appropriate environmental conditions for the fish.
During the 12 weeks experimental period of the
present study, water quality parameters were within
the acceptable range for freshwater fish culture as
indicated by Boyd (1990) and Delince (1992) making
the stocking density as the main variable affecting the
hematological and biochemical parameters of the
studied fish.
The physiological system of fish can be
severely challenged by a variety of biological,
chemical and physical factors. When the
physiological tolerance limits of fish are surpassed,
reproductive success, growth, activity, resistance to
infectious diseases and survival can all be impaired.
Stress elicits a generalized endocrine response in fish
which in turn induces a suite of secondary effects
including rapid mobilization of energy reserves. The
secondary responses of fish to the stressors can be
evaluated by measurement of secondary biochemical
indicators, such as changes in hematology and
plasma chemistry (Wedemeyer et al., 1990).
Fish hematology is gaining great attention in fish
culture because of its importance in monitoring the
health status of fish (Hrubec et al., 2000). In case of
cultured species, occasionally health issues arise that
necessitate clinical evaluation of the fish under such
captive conditions. Lack of published species-specific
normal reference ranges remains the primary reason that
blood testing is not routinely performed in fish health
evaluations (Mauel et al., 2007). The knowledge of the
hematological characteristics is an important tool that
can be used as an effective and sensitive index to
monitor physiological and pathological changes in fish
(Kori-Siakpere et al., 2005).
Alteration of fish blood biochemistry may be
indicative of unsuitable environmental conditions or
the presence of stressing factors (Barcellos et al.,
2004). Our results showed significant higher values
of RBCs, Hb and Ht in the statistical evaluation of T
1

compared to the other two treatments which indicates
that increasing the stocking density over 3 fish/m
3
affected consequently these relevant physiological
parameters of fish. This is confirmed by the
significant decrease in values of RBCs, Hb, Ht and
MCV at stocking density of 6 fish/m
3
(T
2
) followed
by 9 fish/m
3
(T
3
). Similar results have been reported
by Kopp et al. (2010) who studied the influence of
cyanobacterial water bloom on hematological and
biochemical indices of two years old silver carp, H.
molitrix although they reported higher values of
normal RBCs, Hb and Ht than the present study
which may be attributed to the age difference of the
studied fish and changes of the experimental
conditions.
According to Coles (1986) the calculated blood
indices have particular importance in describing
anemia in most animals. The detected non-significant
differences in values of MCH and MCHC support the
assumption that increasing the stocking density did
not pose much challenge to the erythrocytes and did
not indicate any pathological condition in the studied
fish, even if the decrease is significant in values of
RBCs, Hb and Ht separately. This is also confirmed
by the recorded non-significant difference among
values of WBCs. According to Weber and Jensen
(1988) erythropoiesis and hemoglobin synthesis
require long time to complete and can only be
involved in long-term adaptation.
Blood glucose level has been used as an
indicator of environmental stress to reect changes in
carbohydrate metabolism under stress conditions
(Kavitha et al., 2010). Tintos et al. (2006) reported
significant increase in plasma glucose of immature
gilthead sea bream, Sparus auratus after short-term
and long-term stress conditions. This strongly
supports the present findings with the detected highly
significant increase in values of plasma glucose in T
2

and T
3
which were much higher than the normal
reference interval of cyprinids (63-86 mg/dl)
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584

indicated by Nicula et al. (2010) and T
1
showed the
optimum condition regarding this parameter.
The measurement of total protein, albumin and
globulin in serum or plasma is of considerable
diagnostic value in fish as it relates to general
nutritional status as well as the integrity of the vascular
system and liver functions (Abdel-Tawwab et al.,
2008). According to Firat and Kargin (2010) the stress
response of Nile tilapia, Oreochromis niloticus
exposed to individual and combined mixtures of heavy
metals was expressed by significant increase in
glucose, total protein, albumin, AST and ALT as well
as a significant decrease in cholesterol levels. This
strongly supports the present findings where the
significant increase in values of total protein, albumin
and globulin was associated with increasing the
stocking density of silver carp. However, the detected
values of total protein were within the normal
physiological range of cyprinids (2.10-5.76 g/dl) while
values of albumin in case of T
2
and T
3
were marginally
higher than the normal range of cyprinids (0.53-2.20
g/dl) indicated by Nicula et al. (2010).
Cholesterol is one of the structural components
of cell membranes as well as the outer layer of
plasma lipoproteins and is the precursor of all steroid
hormones (Yang and Chen, 2003). Triglycerides
function primarily in providing cellular energy and
can be used as an indicator of nutritional status. In
fact the greatest change in body composition is
usually produced in the lipid fraction (Wallaert and
Babin, 1994). The measured values of cholesterol and
triglycerides showed highly significant decrease
among treatments. Similar trend in cholesterol values
of stressed fish was described by Zhang et al. (2008)
in crucian carp, Carassius auratus and by Qiu et al.
(2009) in silver carp, H. molitrix and bighead carp,
Aristichthys nobilis. During sexual maturation lipid
stores are mobilized and directioned to the gonads to
sustain their development (Guijarro et al., 2003).
This can support the present findings in T
1
and T
2

where the silver carp in this premature developmental
stage had higher levels of cholesterol and
triglycerides in preparation to this shift of body stores
of lipid. The negative effect of stressors on fish may
be indicated by a decrease in body lipid content
(Svobodova et al., 2006). The detected values of
cholesterol in T
1
and T
2
were within the normal
physiological range of cyprinids (65-264 mg/dl)
while it was lower than this range in case of T
3
which
may be attributed to the increased metabolic rate and
decreased food availability. Meanwhile, all measured
values of triglycerides were within the normal range
of cyprinids (68-200 mg/dl) indicated by Nicula et al.
(2010).
AST and ALT are frequently used in the
diagnosis of damage caused by pollutants in various
tissues such as liver, muscles and gills (De la Tore et
al., 2000). It is generally accepted that increased
activity of these enzymes in extracellular uid or
plasma is a sensitive indicator of even minor cellular
damage in the liver of fish (Palanivelu et al., 2005).
AST and ALT are quantitatively important in
transamination of amino acids thereby allowing
interplay between carbohydrate and protein
metabolism during the fluctuating energy demands of
the organism in various adaptive situations (Verma et
al., 1981). Moreover, Palikova et al. (2010) stated
that an increased value of alkaline phosphatase
indicates incorrect secretion of bile due to lower food
intake which may be associated with higher stress.
This coincides with the recorded increase in values of
glucose, total protein, ALP, AST and ALT with
increasing the stocking density of silver carp in the
present study. Acclimation of fish to different
environmental factors results in increased metabolic
activity correlated with changes in the quality and
quantity of certain enzymes involved in energy
metabolism and with compensating modifications in
the rate of protein synthesis (Wallaert and Babin,
1994).
AST and ALT are the most frequently tested
enzymes in fish for indication of cyanobacterial
toxicity. The acute toxicity of microcystins (known
cyanobacterial toxin) is unlikely to occur in silver carp
and chronic exposure to cyanobacteria will not be
detected by changes in ALP, AST and ALT in the
blood plasma (Kopp et al., 2011). This supports our
assumption that stocking density was the main variable
affecting these parameters in the present study. The
detected steady increase in values of ALP, AST and
ALT with increasing the stocking density of silver
carp, even if this increase was non-significant in some
cases, indicates the consistent stress effect of
increasing the stocking density on liver functions of
the fish.
Nicula et al. (2010) indicated acceptable
reference intervals of cyprinids as 46-83 U/l, 26-54
U/l and 9-23 U/l for ALP, AST and ALT
respectively. Generally, values of ALP in T
2
and T
3

only were marginally higher than normal values
while values of AST in the three treatments were
lower than normal. The values of ALT were normal
in T
1
and T
2
but marginally higher in T
3
. According
to Kopp et al. (2011) small differences among
biochemical indices in a fish population are
considered normal.
Plasma creatinine and uric acid levels can be
used as rough indicators of glomerular filtration rate
and kidney functions (Maita et al., 1984). Low levels
of creatinine and uric acid have no significance but
increasing their values indicates several disturbances
in renal system (Maxine and Benjamin, 1985).
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585

Increased level of creatinine is an indicator of kidney
damage, muscular dystrophia and physical exertion
of organisms (Masopust, 1998). Our results of total
protein, ALP, AST, ALT, creatinine and uric acid
support the assumption that the liver and muscle
tissues of silver carp were markedly affected with
increasing the stocking density even with the detected
non-significant increase in values of creatinine.
According to Nicula et al. (2010) the normal
physiological ranges of creatinine and uric acid in
cyprinids are 0.03-0.29 mg/dl and 0.78-2.20 mg/dl
respectively. The detected values of creatinine and
uric acid in T
1
were marginally within the upper
limits of these physiological ranges while T
2
and T
3

values were higher than the normal physiological
ranges of both parameters.
In conclusion, the most optimum condition was
detected in T
1
(3 fish/m
3
) where most of the studied
hematological and biochemical parameters were
essentially normal and within the range consistent
with good fish health. Moreover, measurement of
RBCs count, Hb, Ht, plasma glucose, triglycerides,
total protein, albumin, ALP, AST and uric acid
proved to be more sensitive and reliable in the
detection of even minor stress response of fish.

Corresponding author
Wael A. Omar
Department of Zoology, Faculty of Science, Cairo
University, Egypt
ecowael@yahoo.com

References:
Abdel-Tawwab, M.; Abdel-Rahman A.M. and Ismael, N.E.M.
(2008). Evaluation of commercial live bakers yeast,
Saccharomyces cerevisiae as a growth and immunity
promoter for fry Nile tilapia, Oreochromis niloticus (L.)
challenged in situ with Aeromonas hydrophila. Aquaculture,
280: 185-189.
Allain, C.C.; Poon, L.S.; Chan, C.; Richmond, W. and Fu, P.C.
(1974). Enzymatic determination of total serum cholesterol.
Clin. Chem., 20(4): 470-475.
Barcellos, L.J.G.; Kreutz, L.C.; de Souza, C.; Rodrigues, L.B.;
Fioreze, I.; Quevedo, R.M.; Cericato, L.; Soso, A.B.; Fagundes,
M.; Conrad, J., Lacerda, L.A. and Terra, S. (2004).
Hematological changes in jundia (Rhamdia quelen Quoy and
Gaimard Pimelodidae) after acute and chronic stress caused by
usual aquacultural management, with emphasis on
immunosuppressive effects. Aquaculture, 237: 229-236.
Barham, D. and Trinder, P. (1972). Enzymatic determination
of uric acid. Analyst., 97: 142-145.
Bergmeyer, H.U. (1974). Methods of enzymatic analysis.
Academic Press, New York and London. Vol.4. pp.727-771.
Blaxhall, P.C. and Daisley, K.W. (1973). Routine
haematological methods for use with fish blood. J. Fish
Biol., 5: 771-781.
Boyd, C.E. (1990). Water quality in ponds for aquaculture.
Birmingham Publishing Co. Birmingham, Alabama, USA.
pp.135-170.
Bradford, M.M. (1976). A rapid and sensitive method for the
quantitation of microgram quantities of protein utilizing the
principle of protein-dye binding. Anal. Biochem. 72: 248-
254.
Chen, C.; Wooster, G.A. and Bowser, P.R. (2004).
Comparative blood chemistry and histopathology of tilapia
infected with Vibrio vulnificus or Streptococcus iniae or
exposed to carbon tetrachloride, gentamicin, or copper
sulfate. Aquaculture, 239: 421-443.
Coles, E.H. (1986). Veterinary clinical pathology.
Philadelphia, W.B. Saunders. pp.10-42.
De la Tore, F.R.; Salibian, A. and Ferrari, L. (2000).
Biomarkers assessment in juvenile Cyprinus carpio exposed
to waterborne cadmium. Environ. Pollut., 109: 227-278.
Delince, G. (1992). The ecology of fish pond ecosystem with
special reference of Africa. Kluwer Academic Puplishers,
London. pp.230.
Dumas, B.T. and Biggs, H.G. (1972). Standard methods of
clinical chemistry. Vol. 7. Academic Press, New York, USA.
pp.175.
FAO (Food and Agriculture Organization) (2005).
Hypophthalmichthys molitrix, Cultured Aquatic Species
Information Programme. Text by Yang, N. In: FAO
Fisheries and Aquaculture Department [online report].
Rome. Updated 26 July 2005.
http://www.fao.org/fishery/culturedspecies/Hypophthalmicht
hys_molitrix/en.
Firat, O. and Kargin, F. (2010). Individual and combined
effects of heavy metals on serum biochemistry of Nile tilapia
Oreochromis niloticus. Arch. Environ. Contam. Toxicol., 58:
151-157.
GFCM (General Fisheries Commission for the Mediterranean)
(2010). Synthesis of Mediterranean marine finfish
aquaculture - A marketing and promotion strategy. Studies
and reviews. No.88, Rome, FAO.
Guijarro, A.I.; Lopez-Patino, M.A.; Pinillos, M.L.; Isorna, E.;
De Pedro, N.; Alonso-Gomez, A.L.; Alonso-Bedate, M. and
Delgado, M.J. (2003). Seasonal changes in haematology and
metabolic resources in the tench. J. Fish Biol., 62: 803-815.
Henry, R.J.; Cannon, D.C. and Winkelman, J.W. (1974).
Clinical chemistry, principles and techniques. 2
nd
edition,
Harper and Row. New York, USA. pp.541-553.
Hrubec, T.C.; Cardinale, J.L. and Smith, S.A. (2000).
Hematology and plasma chemistry reference intervals for
cultured Tilapia (Oreochromis hybrid). Vet. Clin. Pathol.,
29: 7-12.
Kavitha, C.; Malarvizhi, A.; Kumaran, S.S. and Ramesh, M.
(2010). Toxicological effects of arsenate exposure on
hematological, biochemical and liver transaminases activity
in an Indian major carp, Catla catla. Food Chem. Toxicol.,
48: 2848-2854.
Kebus, M.J.; Collins, M.T. and Brownfield, M.S. (1992).
Effects of rearing density on the stress response and growth
of rainbow trout. J. Aquat. Anim. Health., 4: 1-6.
Koedprang, W.; Nakajima, M.; Maita, M. and Taniguchi, N.
(2002). Correlation of hematology and plasma chemistry
levels in silver crucian carp Carassius langsdorfii. Fisheries
Sci., 68(4): 721-728.
Kopp, R.; Mares, J.; Palikova, M.; Navratil, S.; Kubicek, Z.;
Zikova, A.; Hlavkova, J. and Blaha, L. (2009). Biochemical
parameters of blood plasma and content of microcystins in
tissues of common carp (Cyprinus carpio L.) from a
hypertrophic pond with cyanobacterial water bloom.
Aquacult. Res., 40: 1683-1693.
Kopp, R.; Palikova, M.; Mares, J.; Navratil, S.; Kubicek, Z.;
and Zikova, A. (2011). Haematological indices are
Life Science Journal, 2011;8(4) http://www.lifesciencesite.com
586

modulated in juvenile carp, Cyprinus carpio L., exposed to
microcystins produced by cyanobacterial water bloom. J.
Fish Dis., 34: 103-114.
Kopp, R.; Palikova, M.; Navratil, S.; Kubicek, Z.; Zikova, A.
and Mares, J. (2010). Modulation of biochemical and
haematological indices of silver carp (Hypophthalmichthys
molitrix Val.) exposed to toxic cyanobacterial water bloom.
Acta Vet. Brno., 79: 135-146.
Kori-Siakpere, O.; Ake, J.E.G. and Idoge, E. (2005).
Haematological characteristics of the African snakehead,
Parachanna obscura. Afr. J. Biotechnol., 4(6): 527-530.
Maita, M.; Shiomitsu, K. and Ikeda, Y. (1984). Health assessment
by the climogram of hemochemical constituents in cultured
yellow tail. Bull. Jap. Soc. Scient., 51: 205-211.
Masopust, J. (1998). Clinical biochemistry. Demanding and
evaluation of biochemical investigation. Karolinum Praha.
pp.832.
Mauel, M.J.; Miller, D.L. and Merrill, A.L. (2007).
Hematologic and plasma biochemical values of healthy
hybrid tilapia (Oreochromis aureus x Oreochromis nilotica)
maintained in a recirculating system. J. Zoo Wildlife Med.,
38(3): 420-424.
Maxine, M. and Benjamin, B.S. (1985). Outline of veterinary
clinical pathology. 3
rd
edition, Colorado State Univ., Printed
in India at Rekha printers PVT. LTD., New Delhi-110020.
McGowan, M.W.; Artiss, J.D.; Strandbergh, D.R. and Zork, B.
(1983). A peroxidase-coupled method for the colorimetric
determination of serum triglycerides. Clin. Chem., 29(3):
538-542.
Misaila, E.R.; Misaila, C.; Artenie, V. and Simalcsik, F.
(2005). Effect of the chronic stress on some parameters of
the metabolic-blood profile (MBP) of the farming
Cyprinides. Fisheries and Aquaculture Development, XXX,
HAKI, Hungary. pp.147-153.
Nicula, M.; Bura, M.; Simiz, E.; Banatean-Dunea, I.; Patruica,
S.; Marcu, A.; Lunca, M. and Szelei, Z. (2010). Researches
concerning reference values assessment of serum
biochemical parameters in some fish species from
Acipenseridae, Cyprinidae, Esocidae and Salmonidae
family. Anim. Sci. Biotech., 43(1): 498-505.
Palanivelu, V.; Vijayavel, K.; Ezhilarasibalasubramanian, S.
and Balasubramanian, M.P. (2005). Influence of insecticidal
derivative (Cartap hydrochloride) from the marine
polychaete on certain enzyme systems of the freshwater fish
Oreochromis mossambicus. J. Environ. Biol., 26: 191-196.
Palikova, M.; Kopp, R.; Mares, J.; Navratil, S.; Kubicek, Z.;
Chmelar, L.; Bandouchova, H. and Pikula, J. (2010).
Selected haematological and biochemical indices of Nile
tilapia (Oreochromis niloticus) reared in the environment
with cyanobacterial water bloom. Acta Vet. Brno., 79: 63-71.
Qiu, T.; Xie, P.; Guo, L. and Zhang, D. (2009). Plasma
biochemical responses of the plaktivorous filter-feeding
silver carp (Hypophthalmichthys molitrix) and bighead carp
(Aristichthys nobilis) to prolonged toxic cyanobacterial
blooms in natural water. Environ. Toxicol. Pharmacol., 27:
350-357.
Reitman, S. and Frankel, S.A. (1957). A colorimetric method
for the determination of serum glutamic oxaloacetic and
glutamic pyruvic transaminases. Am. J. Clin. Pathol., 28(1):
56-63.
Roche, H. and Boge, G. (1996). Fish blood parameters as a
potential tool for identification of stress caused by
environmental factors and chemical intoxication. Mar.
Environ. Res., 41: 27-43.
SAS (Statistical Analysis System) (2006). SAS/STAT Users
Guide, Version 9.1, SAS Institute Inc., Cary, NC, USA.
Shen, J. (2009). A one-stop reference source for
comprehensive, up-to-date information on biology of silver,
largescale silver and bighead carps and environmental risk
assessment. Environ. Biol. Fish., 84: 341-344.
Svobodova, Z.; Vykusova, B.; Modra, H.; Jarkovsky, J. and
Smutna, M. (2006). Haematological and biochemical profile
of harvest-size carp during harvest and post-harvest storage.
Aquacult. Res., 37: 959-965.
Tintos, A.; Miguez, J.M.; Mancera, J.M. and Soengas, J.L.
(2006). Development of a microtitre plate indirect ELISA for
measuring cortisol in teleosts and evaluation of stress
responses in rainbow trout and gilthead sea bream. J. Fish
Biol., 68: 251-263.
Trinder, P. (1969). Determination of glucose in blood using
glucose oxidase with an alternative oxygen acceptor. Ann.
Clin. Biochem., 6: 24-27.
Verma, S.R.; Rani, S. and Delela, R.C. (1981). Isolated and
combined effects of pesticides on serum transaminases in
Mystus vittatus (African catfish). Toxicol. Lett. 8: 67-71.
Wallaert, C. and Babin, P.J. (1994). Effects of temperature
variations on dietary lipid absorption and plasma lipoprotein
concentrations in trout (Oncorhynchus mykiss). Comp.
Biochem. Physiol., 109(B): 473-487.
Weber, R.E. and Jensen, F.B. (1988). Functional adaptations in
hemoglobins from ectothermic vertebrates. Ann. Rev.
Physiol., 50: 161-179.
Wedemeyer, G.A.; Barton, B.A. and McLeay, D.J. (1990).
Stress and acclimation. In: Methods for fish biology.
Schreck, C.C. and Moyle, P.B. (eds). American Fisheries
Society, Bethesda, MD. pp.415-489.
Yang, J.L. and Chen, H.C. (2003). Effects of gallium on
common carp (Cyprinus carpio): Acute test, serum
biochemistry, and erythrocyte morphology. Chemosphere,
53: 877-882.
Zhang, X.; Xie, P.; Wang, W.; Li, D. and Shi, Z. (2008).
Plasma biochemical responses of the omnivorous crucian
carp (Carassius auratus) to crude cyanobacterial extracts.
Fish Physiol. Biochem., 34: 323-329.


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